CN112881684A - Kit for predicting immunotherapy effect - Google Patents

Kit for predicting immunotherapy effect Download PDF

Info

Publication number
CN112881684A
CN112881684A CN202110063031.1A CN202110063031A CN112881684A CN 112881684 A CN112881684 A CN 112881684A CN 202110063031 A CN202110063031 A CN 202110063031A CN 112881684 A CN112881684 A CN 112881684A
Authority
CN
China
Prior art keywords
kit
effect
predicting
immunotherapy
whole blood
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202110063031.1A
Other languages
Chinese (zh)
Inventor
严勇攀
刘忠瑞
王亚娟
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Beijing Gencde Diagnostics Laboratory Co ltd
Original Assignee
Beijing Gencde Diagnostics Laboratory Co ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Beijing Gencde Diagnostics Laboratory Co ltd filed Critical Beijing Gencde Diagnostics Laboratory Co ltd
Priority to CN202110063031.1A priority Critical patent/CN112881684A/en
Publication of CN112881684A publication Critical patent/CN112881684A/en
Pending legal-status Critical Current

Links

Images

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56966—Animal cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/5302—Apparatus specially adapted for immunological test procedures
    • G01N33/5304—Reaction vessels, e.g. agglutination plates
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/70503—Immunoglobulin superfamily, e.g. VCAMs, PECAM, LFA-3
    • G01N2333/70517—CD8
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/70503—Immunoglobulin superfamily, e.g. VCAMs, PECAM, LFA-3
    • G01N2333/70521—CD28, CD152
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/70589—CD45
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/70596—Molecules with a "CD"-designation not provided for elsewhere in G01N2333/705
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Urology & Nephrology (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Hematology (AREA)
  • Cell Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Biochemistry (AREA)
  • Pathology (AREA)
  • Food Science & Technology (AREA)
  • Biotechnology (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Microbiology (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Zoology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Virology (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

The invention discloses a kit for predicting an immunotherapy effect, and particularly relates to the technical field of kits. According to the invention, target cells in peripheral blood mononuclear cells are directly detected, so that a special operation is not required for collecting tumor tissues in a patient body, adverse risks brought to the patient by adopting a method of surgical collection are avoided, the whole operation process is convenient and rapid, and great pain brought to the patient in the process of surgical collection can be effectively reduced, so that the patient and family members thereof can easily accept the tumor tissues, and the blocking agent blocks stimulation of PD-1 and TCR to cause reversion of PD-1 resistance, so that the drug resistance of PD-1 therapy is enhanced, and the PD-1 therapy can present a good treatment effect, so that the method has a wide market, and the predictive biomarker in the invention can benefit the patient treated by PD-1.

Description

Kit for predicting immunotherapy effect
Technical Field
The invention relates to the technical field of kits, in particular to a kit for predicting immunotherapy effect.
Background
In the field of cancer treatment, the worldwide problem of cancer can be overcome in the future, immunotherapy is one of the feasible medical means, and refers to a series of processes capable of enhancing the immune system, inducing or recovering the functions of cytotoxic T cells or other immune effector cells and removing tumor cells, wherein a PD-1 antibody is an immune checkpoint inhibitor, and is a novel drug for tumor immunotherapy, and the PD-1 antibody becomes the most important breakthrough in cancer treatment.
The PD-1 immune checkpoint inhibitor is a novel treatment method, which is guiding a new cancer treatment revolution, and with the two PD-1 monoclonal antibody medicines from foreign medicine enterprises in the market in the land, namely Ophiowa (O medicine) of Poison's, Behcet's and Kreida (K medicine) of Moshadong, PD-1 immune treatment becomes more and more accepted by people and becomes popular cancer treatment means, however, the effects presented by immune treatment are definitely different, some PD-1 immune treatments do not obtain good treatment effects, and some are opposite, which indicates that PD-1 immune treatment can only show better curative effect in a part of patients, and how to accurately find the effective part of patients is a problem of the current clinical treatment, and no method capable of predicting the treatment effect of the patients exists in the market, moreover, in order to distinguish the reagent kit in the prior art conveniently, a user generally needs to label the reagent kit by using a recording tool and then paste the reagent kit on the surface of the reagent kit, which is troublesome, and needs to purchase a writing and pasting tool additionally.
The research patent makes up the defects of predicting the treatment effect, detecting PD1+ CD38+ CD8+ cells by the flow cytometry technology and making up the prediction and therapeutic biomarkers of anti-PD-1 treatment.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention provides a kit for predicting the immunotherapy effect, and the technical problems to be solved by the invention are as follows: the immunotherapy shows that the effect is different, some PD-1 immunotherapy does not obtain good treatment effect, some are opposite, and it is indicated that PD-1 immunotherapy can only show better curative effect in a part of patients, and how to accurately find the part of effective patients is a difficult problem of current clinical treatment, there is no method for predicting the treatment effect of patients on the market, and the reagent kit in the prior art is convenient to distinguish, generally needs a user to label the part of effective patients with a recording tool and then paste the part of effective patients on the surface of the reagent kit, which is troublesome, and needs to purchase writing and pasting tools additionally.
In order to achieve the purpose, the invention provides the following technical scheme: the kit for predicting the immunotherapy effect comprises a shading layer, wherein a kit is arranged in the shading layer, a thread box cover is connected to the outer surface of the shading layer in a threaded mode, three identification rings are fixedly connected to the outer surface of the shading layer respectively, ten arc-shaped elastic clamping frames are fixedly connected to the front sides of the identification rings, the front sides of the shading layer are fixedly connected with the back sides of the three arc-shaped elastic clamping frames, identification rod structures are arranged in the three arc-shaped elastic clamping frames below the shading layer, an antibody reagent combination is arranged in the kit, and the antibody reagent combination comprises APC-CD38, PE-CD45, PECY7-PD-1, AmCYan-CD8 and APC-CD 38.
Use of a kit for predicting the effect of an immunotherapy comprising the steps of:
s1, collecting blood before operation for a patient planned to receive PD-1 immunotherapy, collecting not less than 500ul of EDTA anticoagulated whole blood sample by adopting an EDTA anticoagulation tube, and storing the sample in an environment of 4-10 ℃.
S2, taking APC-CD38, PE-CD45, PECY7-PD-1 and AmCYan-CD8 labeled antibodies 5ul/test respectively on ice in a dark place, and taking APC-CD38 labeled antibodies 3 ul/test.
S3, and slowly inverting the kit 10 times to facilitate uniform mixing of the EDTA-anticoagulated whole blood sample.
S4, adding 100ul of whole blood sample into the flow tube, slightly oscillating for two seconds by using the oscillator, slightly oscillating for two seconds again after one second, circulating for 5 times, and uniformly mixing the whole blood sample.
S5, adding 1ml of xBD Pharm Lyse TM into each tube, performing vortex oscillation and uniform mixing, standing for 10 minutes in a dark place at room temperature, performing centrifugation at 1500 rpm for 5 minutes, pouring out the supernatant after the centrifugation is finished, adding 1ml of PBS into each tube, performing vortex oscillation and uniform mixing, performing centrifugation at 1500 rpm for 5 minutes again, and pouring out the supernatant after the centrifugation is finished.
S6, adding 0.3ml PBS to each tube, keeping away from light at 4 ℃, detecting in 3h, analyzing data according to the data to obtain the content value of endothelial cells, comparing the content value with a defined value, if the relative expression level of the endothelial cells is lower than the defined value, predicting that the immunotherapy effect is effective, and then classifying by the level of effect, difference, poor, good and good, and verifying the predictive biomarker effect.
As a further scheme of the invention: the EDTA-anticoagulated whole blood sample collected in S1 needs to be submitted within 12 hours.
As a further scheme of the invention: the EDTA anticoagulated whole blood sample collected in S1 should be detected within 24 hours after the sample is taken.
As a further scheme of the invention: the EDTA anticoagulated whole blood sample collected in S1 is prohibited from being frozen throughout the testing process.
As a further scheme of the invention: the whole blood specimen in S4 needs to be kept under dark conditions at room temperature for 20 minutes after being mixed uniformly.
As a further scheme of the invention: the identification rod structure comprises a support rod, and a rubber layer is arranged on the outer surface of the support rod.
As a further scheme of the invention: the lower surface of the shading layer is fixedly connected with a plurality of rubber particles.
As a further scheme of the invention: the left side surface and the right side surface of the threaded box cover are respectively provided with two anti-skidding bulges.
The invention has the beneficial effects that:
1. according to the invention, target cells in peripheral blood mononuclear cells are directly detected, so that a special operation is not required for collecting tumor tissues in a patient body, adverse risks brought to the patient by adopting a method of surgical collection are avoided, the whole operation process is convenient and rapid, and great pain brought to the patient in the process of surgical collection can be effectively reduced, so that the patient and family members thereof can easily accept the tumor tissues, and the blocking agent blocks stimulation of PD-1 and TCR to cause reversion of PD-1 resistance, so that the drug resistance of PD-1 therapy is enhanced, and the PD-1 therapy can present a good treatment effect, so that the method has a wide market, and the predictive biomarker in the invention can benefit the patient treated by PD-1.
2. The invention sets the arc elastic clamping frame and the identification rod structure, when the whole reagent box needs to be labeled, a user can take down the identification rod structure according to the size of the label, the first layer of identification ring is arranged in hundreds, the second layer of identification ring is arranged in tens, the third layer of identification ring is arranged in units, when the label is 123, the first identification rod structure is clamped into the first arc elastic clamping frame in the first layer of identification ring, then the second identification rod structure is clamped into the second arc elastic clamping frame in the second layer of identification ring, and then the third identification rod structure is clamped into the third arc elastic clamping frame in the third layer of identification ring, so that the invention does not need to label different reagent boxes and then paste the reagent boxes on the surface without additionally purchasing a writing and pasting tool, is simple and convenient, and the device can be repeatedly used.
3. The light shielding layer is arranged on the surface of the kit, so that the light shielding layer can perform light shielding treatment on APC-CD38, PE-CD45, PECY7-PD-1, AmCYan-CD8 and APC-CD38 in the kit, and the light shielding use of the kit is facilitated, and the bottom of the light shielding layer is provided with a plurality of rubber particles, and the rubber material has certain skid resistance, so that the device can have certain skid resistance effect when placed on a desktop, is not easy to slip due to small force, ensures the placing stability of the kit, and the rubber layer is arranged on the surface of the support rod, so that the support rod can be clamped in the arc-shaped elastic clamping frame more firmly due to the excellent skid resistance of the rubber layer, and is not easy to fall off, and the fluorescent material is arranged in the rubber layer, so that a user can clearly see the label of the kit even under a dim condition, so that the use effect is ideal.
Drawings
FIG. 1 is a front view of the present invention;
FIG. 2 is a schematic top view of the present invention;
FIG. 3 is a schematic top view of the arc-shaped elastic clamping frame of the present invention;
FIG. 4 is a schematic cross-sectional view of a recognition rod according to the present invention;
FIG. 5 is a graph showing statistical broken lines of different results;
FIG. 6 is a graph showing the results of a test performed on a patient with better clinical performance after PD-1 treatment;
FIG. 7 is a graph showing the results of a test for poor clinical performance of PD-1 in a patient;
in the figure: 1 light shield layer, 2 kit, 3 screwed cap, 4 discernment circles, 5 arc elasticity card framves, 6 discernment pole structures, 61 bracing pieces, 62 rubber layer, 7 rubber granules.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
As shown in fig. 1-7, the present invention provides a kit for predicting immunotherapy effect, which comprises a light shielding layer 1, wherein the light shielding layer 1 is arranged, such that APC-CD38, PE-CD45, PECY7-PD-1, AmCYan-CD8, and APC-CD38 can be conveniently used in a dark place, a kit 2 is arranged in the light shielding layer 1, a screw cap 3 is screwed on an outer surface of the light shielding layer 1, three identification rings 4 are respectively and fixedly connected on an outer surface of the light shielding layer 1, by arranging the three identification rings 4, a user can arrange the first identification ring 4 in a hundred place, arrange the second identification ring 4 in a ten place, arrange the third identification ring 4 in a unit place, when the mark is 123, the user only needs to clip the first identification rod structure 6 into the first arc-shaped elastic clip frame 5 in the first identification ring 4, then clip the second identification rod structure 6 into the second arc-shaped elastic clip frame 5 in the second identification ring 4, then the third identification rod structure 6 is clamped into a third arc-shaped elastic clamping frame 5 in a third layer of identification ring 4, ten arc-shaped elastic clamping frames 5 are fixedly connected with the front surface of the identification ring 4, the front surface of the shading layer 1 is fixedly connected with the back surfaces of the three arc-shaped elastic clamping frames 5, by arranging the three arc-shaped elastic clamping frames 5 on the front surface of the shading layer 1, the three identification rod structures 6 can be respectively clamped into the three arc-shaped elastic clamping frames 5 on the front surface of the shading layer 1 when the identification rod structures 6 are not needed, so that a user can recognize that identification rod structures 6 are arranged in the three lower arc-shaped elastic clamping frames 5 in the kit 2 without the marks, and the kit 2 is internally provided with antibody reagent combinations of APC-CD38, PE-CD45, PECY7-PD-1, AmCYan-CD8 and APC-CD 38.
Use of a kit for predicting the effect of an immunotherapy comprising the steps of:
s1, collecting blood before operation for a patient planned to receive PD-1 immunotherapy, collecting not less than 500ul of EDTA anticoagulated whole blood sample by adopting an EDTA anticoagulation tube, and storing the sample in an environment of 4-10 ℃.
S2, taking APC-CD38, PE-CD45, PECY7-PD-1 and AmCYan-CD8 labeled antibodies 5ul/test respectively on ice in a dark place, and taking APC-CD38 labeled antibodies 3 ul/test.
S3, and slowly inverting the kit 2 10 times to facilitate uniform mixing of the EDTA-anticoagulated whole blood sample.
S4, adding 100ul of whole blood sample into the flow tube, slightly oscillating for two seconds by using the oscillator, slightly oscillating for two seconds again after one second, circulating for 5 times, and uniformly mixing the whole blood sample.
S5, adding 1ml of xBD Pharm Lyse TM into each tube, performing vortex oscillation and uniform mixing, standing for 10 minutes in a dark place at room temperature, performing centrifugation at 1500 rpm for 5 minutes, pouring out the supernatant after the centrifugation is finished, adding 1ml of PBS into each tube, performing vortex oscillation and uniform mixing, performing centrifugation at 1500 rpm for 5 minutes again, and pouring out the supernatant after the centrifugation is finished.
S6, adding 0.3ml PBS to each tube, keeping away from light at 4 ℃, detecting in 3h, analyzing data according to the data to obtain the content value of endothelial cells, comparing the content value with a defined value, if the relative expression level of the endothelial cells is lower than the defined value, predicting that the immunotherapy effect is effective, and then classifying by the level of effect, difference, poor, good and good, and verifying the predictive biomarker effect.
The EDTA-anticoagulated whole blood sample collected in S1 needs to be submitted for testing within 12 hours.
EDTA anticoagulated whole blood samples collected in S1 should be assayed within 24 hours after sample collection.
The EDTA anticoagulated whole blood sample collected in S1 was not frozen throughout the assay.
The whole blood specimen in S4 was left to stand in the dark at room temperature for 20 minutes after being mixed uniformly.
The identification rod structure 6 comprises a support rod 61, a rubber layer 62 is arranged on the outer surface of the support rod 61, and the rubber layer 62 is arranged on the surface of the support rod 61, so that the support rod 61 can be clamped in the arc-shaped elastic clamping frame 5 more firmly and is not easy to fall off due to the excellent anti-skid property of the rubber layer 62, a fluorescent material is arranged in the rubber layer 62, the label of the kit 2 can be clearly seen even if a user is in a dark condition, and the use effect is ideal.
The lower surface fixedly connected with a plurality of rubber granule 7 of light shield layer 1 through set up rubber granule 7 in light shield layer 1 bottom, because rubber materials has certain antiskid nature for this device is placed and can has certain anti-skidding effect when the desktop, and difficult emergence is slided.
The left side surface and the right side surface of the threaded box cover 3 are respectively provided with two anti-skidding bulges, and the left side surface and the right side surface of the threaded box cover 3 are respectively provided with two anti-skidding bulges, so that the friction force between the threaded box cover 3 and a palm of a user can be improved when the user twists the threaded box cover 3, and the rotation of the threaded box cover 3 by the user is more convenient and labor-saving.
Examples
Eighteen patients who planned to receive PD-1 immunotherapy were subjected to preoperative blood collection, and the combination of detection reagents and procedures thereof were carried out by flow cytometry as follows:
s1, collecting not less than 500ul of EDTA anticoagulated whole blood samples by adopting EDTA anticoagulation tubes for twenty patients planned to receive PD-1 immunotherapy respectively, and storing the samples in an environment of 4-10 ℃.
S2, taking APC-CD38, PE-CD45, PECY7-PD-1 and AmCYan-CD8 labeled antibodies 5ul/test respectively on ice in a dark place, and taking APC-CD38 labeled antibodies 3 ul/test.
S3, and slowly inverting the kit 2 10 times to facilitate uniform mixing of the EDTA-anticoagulated whole blood sample.
S4, adding 100ul of whole blood sample into the flow tube, slightly oscillating for two seconds by using the oscillator, slightly oscillating for two seconds again after one second, circulating for 5 times, and uniformly mixing the whole blood sample.
S5, adding 1ml of xBD Pharm Lyse TM into each tube, performing vortex oscillation and uniform mixing, standing for 10 minutes in a dark place at room temperature, performing centrifugation at 1500 rpm for 5 minutes, pouring out the supernatant after the centrifugation is finished, adding 1ml of PBS into each tube, performing vortex oscillation and uniform mixing, performing centrifugation at 1500 rpm for 5 minutes again, and pouring out the supernatant after the centrifugation is finished.
S6, adding 0.3ml PBS to each tube, keeping away from light at 4 ℃, detecting in 3h, analyzing data according to the data to obtain the content value of endothelial cells, comparing the content value with a defined value, if the relative expression level of the endothelial cells is lower than the defined value, predicting that the immunotherapy effect is effective, and then classifying by the level of effect, difference, poor, good and good, and verifying the predictive biomarker effect.
Test results and their corresponding effects were obtained for 1-18 patients scheduled to receive PD-1 immunotherapy, respectively:
example 1: the result is 29.18, and the effect column is poor.
Example 2: the result was 8.25, and the effect was listed as better.
Example 3: the result was 7.65, and the effect was listed as good.
Example 4: the result was 20.6, and the effect was ranked as poor.
Example 5: the result was 19.3, and the effect column was poor.
Example 6: the result was 30, and the effect was classified as poor.
Example 7: the result was 22.1, and the effect column was poor.
Example 8: the result was 9.6, and the effect was listed as good.
Example 9: the result was 10.5, and the effect was listed as good.
Example 10: the result was 9.8, and the effect was listed as good.
Example 11: the result was 22.9, and the effect column was poor.
Example 12: the result was 5.9, and the effect was listed as better.
Example 13: the result was 20.6, and the effect was ranked as poor.
Example 14: the result was 16.7 and the effect column was poor.
Example 15: the result was 21.5, and the effect was classified as poor.
Example 16: the result was 5.9, and the effect was listed as good.
Example 17: the result was 10.0, and the effect column was poor.
Example 18: the result was 9.4, and the effect was listed as good.
Comparative example:
the tissue of the diseased part of the patient is sampled and sliced by adopting an operation mode, and then the immunotherapy effect of the slices is predicted.
In summary, in the comparative example, the patient needs to be sampled by surgery, which is not only more harmful and more troublesome, but also wastes resources, and also causes more pain to the patient, so that the patient and the family members need to consume more money and are difficult to accept, and the test results of PD1+ CD38+ CD8+ in the blood in the example show that the detection value is lower, the immunotherapy effect is good, the detection value is higher, and the immunotherapy effect is poor, in the tests of the eighteen patients, the number of good treatment effects is 2, the number of good treatment effects is 6, the number of poor treatment effects is 4, and the number of poor treatment effects is 6, so that PD1+ CD38+ CD8+ can be used as a predictive and therapeutic biomarker for anti-PD-1 therapy.
The working principle of the invention is as follows: when the whole kit 2 needs to be labeled, a user can take down the lower identification rod structure 6 according to the size of the label, the first layer of identification ring 4 is arranged in hundreds, the second layer of identification ring 4 is arranged in tens, and the third layer of identification ring 4 is arranged in units, when the label is 123, the user only needs to clamp the first identification rod structure 6 into the first arc-shaped elastic clamping frame 5 in the first layer of identification ring 4, then clamp the second identification rod structure 6 into the second arc-shaped elastic clamping frame 5 in the second layer of identification ring 4, and then clamp the third identification rod structure 6 into the third arc-shaped elastic clamping frame 5 in the third layer of identification ring 4, and the invention directly detects target cells in single nuclear cells of peripheral blood, so that the operation is not needed to collect tumor tissues in a patient body, and the adverse risk brought to the patient by the operation collection method is avoided, and blocking of PD-1 and TCR stimulation by the blocking agent can result in reversal of PD-1 resistance, and drug resistance of PD-1 therapy is enhanced, so that PD-1 therapy can present a better therapeutic effect, and the predictive biomarker provided by the invention can benefit patients treated by PD-1.
The points to be finally explained are: first, in the description of the present application, it should be noted that, unless otherwise specified and limited, the terms "mounted," "connected," and "connected" should be understood broadly, and may be a mechanical connection or an electrical connection, or a communication between two elements, and may be a direct connection, and "upper," "lower," "left," and "right" are only used to indicate a relative positional relationship, and when the absolute position of the object to be described is changed, the relative positional relationship may be changed;
secondly, the method comprises the following steps: in the drawings of the disclosed embodiments of the invention, only the structures related to the disclosed embodiments are referred to, other structures can refer to common designs, and the same embodiment and different embodiments of the invention can be combined with each other without conflict;
and finally: the above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that are within the spirit and principle of the present invention are intended to be included in the scope of the present invention.

Claims (9)

1. The kit for predicting the immunotherapy effect is characterized by comprising a shading layer (1), wherein a kit (2) is arranged in the shading layer (1), a screw box cover (3) is connected to the outer surface of the shading layer (1) in a threaded manner, three identification rings (4) are respectively and fixedly connected to the outer surface of the shading layer (1), ten arc-shaped elastic clamping frames (5) are fixedly connected to the front surfaces of the identification rings (4), the front surfaces of the shading layer (1) and the back surfaces of the three arc-shaped elastic clamping frames (5) are fixedly connected, identification rod structures (6) are arranged in the three lower arc-shaped elastic clamping frames (5), and an antibody reagent combination is arranged in the kit (2) and comprises APC-CD38, PE-CD45, PECY7-PD-1, AmCYan-CD8 and APC-CD 38.
2. The use of a kit for predicting the effect of an immunotherapy according to claim 1, characterized by comprising the steps of:
s1, collecting blood before operation for a patient planned to receive PD-1 immunotherapy, collecting not less than 500ul of EDTA (ethylene diamine tetraacetic acid) anticoagulated whole blood sample by adopting an EDTA anticoagulation tube, and storing the sample in an environment of 4-10 ℃;
s2, taking APC-CD38, PE-CD45, PECY7-PD-1 and AmCYan-CD8 labeled antibodies 5ul/test respectively on ice in a dark place, and taking APC-CD38 labeled antibodies 3 ul/test;
s3, slowly inverting the kit (2) for 10 times so as to uniformly mix the EDTA anticoagulated whole blood sample;
s4, adding 100ul of whole blood sample into the flow tube, slightly oscillating for two seconds by using the oscillator, slightly oscillating for two seconds again after one second, circulating for 5 times, and uniformly mixing the whole blood sample;
s5, adding 1ml of multiplied by BD Pharm Lyse TM into each tube, performing vortex oscillation and uniform mixing, standing for 10 minutes in a dark place at room temperature, performing centrifugation at 1500 rpm for 5 minutes, pouring out the supernatant after the centrifugation is finished, adding 1ml of PBS into each tube, performing vortex oscillation and uniform mixing, performing centrifugation at 1500 rpm for 5 minutes, and pouring out the supernatant after the centrifugation is finished;
s6, adding 0.3ml PBS to each tube, keeping away from light at 4 ℃, detecting in 3h, analyzing data according to the data to obtain the content value of endothelial cells, comparing the content value with a defined value, if the relative expression level of the endothelial cells is lower than the defined value, predicting that the immunotherapy effect is effective, and then classifying by the level of effect, difference, poor, good and good, and verifying the predictive biomarker effect.
3. Use of a kit for predicting the effect of an immunotherapy according to claim 2, characterized in that: the EDTA-anticoagulated whole blood sample collected in S1 needs to be submitted within 12 hours.
4. Use of a kit for predicting the effect of an immunotherapy according to claim 2, characterized in that: the EDTA anticoagulated whole blood sample collected in S1 should be detected within 24 hours after the sample is taken.
5. Use of a kit for predicting the effect of an immunotherapy according to claim 2, characterized in that: the EDTA anticoagulated whole blood sample collected in S1 is prohibited from being frozen throughout the testing process.
6. Use of a kit for predicting the effect of an immunotherapy according to claim 2, characterized in that: the whole blood specimen in S4 needs to be kept under dark conditions at room temperature for 20 minutes after being mixed uniformly.
7. Use of a kit for predicting the effect of an immunotherapy according to claim 1, characterized in that: the identification rod structure (6) comprises a support rod (61), and a rubber layer (62) is arranged on the outer surface of the support rod (61).
8. Use of a kit for predicting the effect of an immunotherapy according to claim 1, characterized in that: the lower surface of the shading layer (1) is fixedly connected with a plurality of rubber particles (7).
9. Use of a kit for predicting the effect of an immunotherapy according to claim 1, characterized in that: the left side surface and the right side surface of the screw thread box cover (3) are respectively provided with two anti-skidding bulges.
CN202110063031.1A 2021-01-18 2021-01-18 Kit for predicting immunotherapy effect Pending CN112881684A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202110063031.1A CN112881684A (en) 2021-01-18 2021-01-18 Kit for predicting immunotherapy effect

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202110063031.1A CN112881684A (en) 2021-01-18 2021-01-18 Kit for predicting immunotherapy effect

Publications (1)

Publication Number Publication Date
CN112881684A true CN112881684A (en) 2021-06-01

Family

ID=76049084

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202110063031.1A Pending CN112881684A (en) 2021-01-18 2021-01-18 Kit for predicting immunotherapy effect

Country Status (1)

Country Link
CN (1) CN112881684A (en)

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102502118A (en) * 2011-10-24 2012-06-20 马广忠 Conveniently-identified beverage bottle and application method thereof
US20190336600A1 (en) * 2018-05-01 2019-11-07 Augusta University Research Institute, Inc. Methods for detecting and reversing immune therapy resistance
WO2020056346A1 (en) * 2018-09-13 2020-03-19 The Trustees Of The University Of Pennsylvania Interferon pathway genes regulate and predict efficacy of immunotherapy
CN111344568A (en) * 2017-09-12 2020-06-26 免疫特征私人有限公司 Predicting response to immunotherapy
CN111551733A (en) * 2020-05-29 2020-08-18 武汉大学 Method for quantitatively detecting content of immune cell-derived extracellular vesicle PD-1, ELISA kit and using method
WO2020207771A1 (en) * 2019-04-10 2020-10-15 Universität Zürich A method for determining the likelihood of a patient being responsive to cancer immunotherapy

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102502118A (en) * 2011-10-24 2012-06-20 马广忠 Conveniently-identified beverage bottle and application method thereof
CN111344568A (en) * 2017-09-12 2020-06-26 免疫特征私人有限公司 Predicting response to immunotherapy
US20190336600A1 (en) * 2018-05-01 2019-11-07 Augusta University Research Institute, Inc. Methods for detecting and reversing immune therapy resistance
WO2020056346A1 (en) * 2018-09-13 2020-03-19 The Trustees Of The University Of Pennsylvania Interferon pathway genes regulate and predict efficacy of immunotherapy
WO2020207771A1 (en) * 2019-04-10 2020-10-15 Universität Zürich A method for determining the likelihood of a patient being responsive to cancer immunotherapy
CN111551733A (en) * 2020-05-29 2020-08-18 武汉大学 Method for quantitatively detecting content of immune cell-derived extracellular vesicle PD-1, ELISA kit and using method

Similar Documents

Publication Publication Date Title
JP2022043302A (en) Biodosimetry panels and methods
DANS et al. Inappropriate use of the cerebrospinal fluid Venereal Disease Research Laboratory (VDRL) test to exclude neurosyphilis
CN114460286A (en) Assay for capturing and detecting circulating multiple myeloma from blood
CN205748983U (en) A kind of saliva collecting device and saliva detection kit
CN101403740A (en) Mass spectrum model used for detecting liver cancer characteristic protein and preparation method thereof
Zenouz et al. Relationship between lichen planus and Helicobacter pylori infection
Shin Unexpected red cell antibody detection by conditional combination of LISS/Coombs and NaCl/Enzyme gel tests at a tertiary care hospital in Korea: a 5-year study
CN112881684A (en) Kit for predicting immunotherapy effect
Marchand et al. Optimal collection technique and devices for a quality pap smear
Tan et al. Clinical significance of detecting HLA-DR, 14-3-3η protein and d-dimer in the diagnosis of rheumatoid arthritis
Kalukula et al. Translating cell mechanobiology and nuclear deformations to the clinic
Cui et al. The length and distribution of plasma cell-free DNA fragments in stroke patients
Chen et al. Mobile laboratory in Sierra Leone during outbreak of Ebola: practices and implications
JP6822006B2 (en) Target particle recovery method and recovery device
CN108728400A (en) A kind of Chronic glomerular disease glomerulosclerosis marker and its detection kit
Bustos et al. EVALUATION OF A NEW RAPID DIAGNOSTIC TEST “DETERMINE™ MALARIA PF” AGAINST STANDARD BLOOD FILM, ICT MALARIA PF™ AND PARASIGHT™ F
Rawat et al. Dermatoglyphic Patterns in Breast Cancer Patients
Otoni et al. Chemokine profile in the sera and urine of patients with schistosomal glomerulopathy
CN115165829A (en) Method for quantitatively analyzing immune cells and functional proteins of systemic lupus erythematosus and application of method
Imai et al. Volume-corrected mitotic index as a prognostic factor in oral squamous cell carcinomas
CN218870295U (en) Laboratory mouse blood sample collection device
Niyibizi et al. Research Article Diagnostic Performance between Histidine-Rich Protein 2 (HRP-2), a Rapid Malaria Diagnostic Test and Microscopic-Based Staining Techniques for Diagnosis of Malaria
CN113341118A (en) Method for testing clinical test items
GS et al. Prevalence of Anti-Nuclear Antibody Pattern-One Year Experience in a Tertiary Care Center
Xu et al. Cancer Patient Management Strategy in a Cancer Center of Zhejiang, China During the COVID-19 Pandemic (preprint)

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
RJ01 Rejection of invention patent application after publication

Application publication date: 20210601

RJ01 Rejection of invention patent application after publication